App/. Radiat. hr. Vol. 41, No. 3, pp. 327-331, 1990 Inr. J. Radiat. Appl. Insrrum. Part A Printed in Great Britain. All rights reserved

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Accumulation of Each Element in the

Lysosome of Tumor and Liver Cells as

Systematized According to its Location in

the Periodic Table

ATSUSHI ANDO* and lTSUK0 AND0

School of Allied Medical Professions, Kanazawa University, 5-I l-80, Kodatsuno, Kanazawa 920. Japan

(Receioed 8 May 1989; in revised form 31 July 1989)

Hard and borderline acids which were formed by losing electrons from the d-shell accumulated extensively in the lysosomes of tumor and liver with time after administration of these ions. They were bound in these tissues to acid mucopolysaccharides, the mol. wt of which exceed 40,000 Da. Trivalent hard acids which were formed by losing electrons from s- or p-shells accumulated extensively in liver lysosomes, but very little in tumor lysosomes. and were bound in the tissues to the acid mucopolysaccharides with a mol. wt of ca 10,000 Da.

Introduction

Swartzendruber rt ul. (1971) have shown that intra- cellular 67Ga present in normal and neoplastic tissue is localized in lysosome-like bodies; there have been two conflicting opinions about the lysosomal accu- mulation of 67Ga in the tissues ever since. Some investigators (Aulbert and Haubold, 1974; Takeda et al., 1977) support their view; however, others (Deck- ner et ul., 1971; Orii, 1972; Ando et al., 1982a) claim that the lysosome does not play a major role in the tumor accumulation of 67Ga, while it may play an important role in the liver accumulation of this nuclide. To date we have evaluated the lysosomal role in the accumulation of 19 radioactive metal ions in tumor and liver. We found on the basis of our results and those described by other investigators a very interesting relationship between the location of these elements in the Thomsen-Bohr type periodic table and the lysosomal accumulation of metal ions.

Materials and Methods

Male Donryu rats (body weight 159 k 27 g) under- went S.C. implantation of Yoshida sarcoma (1 x IO4 cells/O.25 mL) in the right thigh. Six to seven days later an appropriate amount of radioactive nuclide was administered, at which time the tumor had grown to 1.5-2.0 cm in diameter.

*Author for correspondence.

Radioactive inorganic compounds in solution were prepared as carrier-free nuclides or as those contain- ing little stable nuclide. Chemical forms and injected doses were described previously (Ando er al., 1985, 1989a). These labeled compounds were injected intra- venously into the rats. Liver and tumor tissue were excised from 10 min to 48 h after the administration of these compounds, and were homogenized sepa- rately. Subcellular fractionation was carried out according to the modified method (Ando et ul., 1982a) of Hogeboom and Schneider.

Results and Discussion

Accumulation and accumulation ratio

When the radioactivities of nuclear fraction, mito- chondrial fraction (lysosomes are contained in this fraction), microsomal fraction and supernatant frac- tion are expressed as A(cpm), B(cpm), C(cpm) and D(cpm), respectively, the accumulation (percentage) of the mitochondrial fraction can be calculated by the following formula:

B

A+B+C+D x lOO(%).

The accumulation ratio was calculated by dividing the 48 h value by the 10 min value.

The accumulation (%) of these nuclides in the mitochondrial fraction (containing lysosomes) of liver is shown in Table 1. The accumulation ratios (48 h/10 min) are also shown in this table. Hard acids

321

ATSUSHI ANDO and ITSUKO ANDO

(Ga”, In’+, Yb3’, Tm”, Cr’+, SC”+. Z?‘, HfJ’, Nb’+. Ta5+) of tri-, quadri- and pentavalence. and some borderline acids (Co’ ‘+, Ru’ ‘+ ) accumulated extensively in this fraction as time passed after ad- ministration. The accumulation ratios for these metal ions were between 2.4 and 6.9, except for Hf4+. It is clear from the studies previously reported that “‘Ga (Brown et al., 1973) and “‘In (Takeda et cd., 1977) accumulated in liver lysosomes. Recently we (Ando et al., 1989b) determined that “6Sc accumulated in tumor and liver lysosomes. We regard it as probable that the elements “‘Yb (Ando et al.. 1981). ““Tm (Ando et al., 1983a), “Cr (Ando et al., 1987a). “Zr, “I Hf (Ando and Ando, 1986). “Nb. “‘Ta (Ando and Ando, 1980), ‘“Co and ‘“2Ru (Ando et al., 1988a) are contained in the above mitochondrial fraction in the same way that h7Ga, “‘In and “6Sc exist in lysosomes of each tissue. We can thus conclude from these results that lysosomes played an important role in their accumulation in the liver. Alkaline metals (Na +. Rb+, Cs+ ), soft acids (Tl’. Hg’+. Pd’+ ) and some (Zn2+) borderline acids accumulated weakly in the mitochondrial fraction of the liver; the accumulation ratios (48 h/l0 min) for these metal ions were all about 1 .O. Lysosomes did not play an important role in the accumulation of these nuclides in the liver. Table 1 also shows the accumulation (%). and the accumulation ratios (48 h/IO min) of all these nuclides in the mitochondrial fraction (containing lysosome) of Yoshida sarcoma. Na’ , Rb’ , Cs .

Zn’+, Tl+, Hg*+ and Pd2+ accumulated weakly in this fraction. In the case of Cr3 +, SC’.+, Zr4 + . Hf4 +. Nb5+ and Ta’+, the accumulation increased markedly from 10 min to 48 h. Accumulation of Co’ ‘+ and Ru’ 4 + also increased with time after administration. These results were similar to those for liver. Conversely, the accumulation of Ga3+, In”+, Yb’+ and Tm’+ hardly increased with time after administration, These results obviously differed from those for liver. Namely, the lysosomal role in the accumulation of these 4 elements in the tissue became weak with the transformation of the tissue into malignant tumor (Ando et al., 1981, 1982a, 1983a).

Location of elements in the Thomsen-Bohr-type

periodic table and accumulation ratio

The accumulation ratios (48 h/IO min) for liver and Yoshida sarcoma are shown below each element in the Thomsen-Bohr-type periodic table (Fig. 1). For elements of the first transition series, the 3d-orbital is filled with electrons from SC to Ni. In the second transition series, the 4d-orbital is filled with electrons from Y to Rh. In the third transition series, the Sd-orbital is filled with electrons from Hf to Pt. SC, Cr and Co have incomplete 3d-shells; Zr, Nb and Ru have incomplete 4d-shells; and Hf and Ta have incomplete Sd-shells. These 8 elements with incom- plete d-shells were transported into lysosomes of the malignant tumor and liver by the acid mucopolysac- charides the mol. wt of which exceeds 40,000 Da. In

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330 ATSUSHI ANDO and ITSUKO ANDO

Table 2. Binding substances (or status) for metal ions m hver and tumor

Metal ion Binding substances (or status)

Gal+, In’- ,Jb’- Tm” % 1 Acid mucopolysaccharide with mol. v,t UI IO.000 Da (trivalent hard acids which were formed by losing electrons from J- or p-shells)

SC”. 0”. Zr’-. Hf”, Nb’-. Ta’+ Acid mucopolysaccharides wth mol. wt exceedmg 40.000 D.1 (Hard acids which were formed by losing electrons from the cl-shell) c’“’ 1-, Ru’ 44 Acid mucopolysaccharides ulth mol. wt rxceedmp JO.000 D.I (borderhne acids which were formed by losing electrons (partly bound) from the (l-shell)

Nn’. Rb-. Cs’. TI’ (mns of alkaline met& and Tl) zn2-, Hg:*. Pd:+

(some borderline and soft acids)

Exists mostly as a free eon

-SH radicals rn protein

addition. trivalent hard acids (Ga’+, In”, Yb”, Tm’+ ) which were formed by losing electrons from S- or p-shells were transported into liver lysosomes by the acid mucopolysaccharide with a mol. wt of ca 10,000 Da. For other elements, only small amounts accumulated into lysosomes of malignant tumors.

Binding substance (or status) ,for metal ions and

l~~sosomul uccumulation

In 1979, Ando originally determined that “Ga, “I In and 16’Yb were bound to the acid mucopolysac- charide in tumor and in liver. It was also reported that “Ga-binding acid mucopolysaccharide could be separated by cellulose acetate electrophoresis from tumor and liver (Ando et al., 1983b). Later, we determined the binding substances for many radio- active metal ions in tumor and liver.

On the chemical bond of metal compounds. the following rules apply (Pearson, 1963). A number of Lewis acids of diverse types are classified as hard acids and soft acids. Hard acids prefer to bind to hard bases. Soft acids prefer to bind to soft bases. Among the metal ions, Gal+, Ini+, Yb’+. Tm3+. Cr”, SC’+, Zr”‘, Hf4’. Nb5+. Ta’+ are hard acids, Hg”. Pd” are soft acids, and CO’+. Zn’+, Ru’+ are borderline cases. Among the body constituents of animals. R-SO; , R-PO:-. and R-COO are hard bases. and R-S -, and R--SH are soft bases. It is reasonable to assume that metallothioneins in tumor tissue are composed of protein and soft acids such as Hg’+. It is also reasonable to assume that hard acids such as Ga’+ and Nb”+ would bind to hard bases such as R-SO?- , R-PO:- and R-COO -.

Substances to which metal ions are bound in tissues are summarized in Table 2. Ga? +, In3+, Yb3+ and Tm’+. which are trivalent hard acids, were bound to the acid mucopolysaccharide with a mol. wt of ca 10,OOODa (Ando et al., 1982b, 1983a. 1983b). We have already reported that the main 67Ga-binding acid mucopolysaccharide was keratan polysulfate or other oversulfated acid mucopolysaccharides (Ando et al., 1987b), and that SC’+, Cr3+, Zr4+, Hf4+, Nb5+ and Ta’+ were bound to the acid mucopoly- saccharides with mol. wt exceeding 40,000 Da (Ando and Ando, 1980, 1986; Ando et al., 1987a). We also

reported that Co’ ’ t and Ru’ ” were partly bound to the acid mucopolysaccharides with mol. wt exceeding 40,000 Da (Ando et al., 1988a). It can be assumed with confidence that these ions which were formed by losing electrons from the d-shell. bind to acid muco- polysaccharides with mol. wt exceeding 40,000 Da. and that they are transported into lysosomes of tumor and liver by these acid mucopolysaccharidcs. It is probable that trivalent hard acids. which were formed by losing electrons from s- or p-shells, bind to the acid mucopolysaccharide with a mol. vit of CCI 10,000 Da. and are thus transported into liver lysosomes.

It is well known that metallothioneins in the liver are composed of -SH radicals in protein and soft acids such as Hg’+ (Winge et ul., 1975). Zn”. Hp” and Pd’+, which were bound to SH radicals, accu- mulated weakly in the mitochondrial fraction of both liver and tumor. Alkaline metals and TI existed mostly as free ions in tissues (Ando et (11.. 1987c, 1988b); they hardly accumulated in lysosomes.

It is highly probable that the difference in lysoso- ma1 accumulation of these ions in tumor and liver is caused by a difference in the binding substances of these ions in the tissues.

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Periodic table and lysosomal accumulation 331

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